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Design of a secondary alcohol degradation pathway from Pseudomonas fluorescens DSM 50106 in an engineered Escherichia coli

  • Applied Genetics and Molecular Biotechnology
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Abstract

The genes encoding an alcohol dehydrogenase, Baeyer–Villiger monooxygenase and an esterase from P. fluorescens DSM 50106, which seemed to be metabolically connected based on the sequence of the corresponding open reading frames, were cloned into one vector (pABE) and functionally expressed in Escherichia coli. Overall expression levels were quite low, however, using whole cells of E. coli JM109 pABE expressing the three recombinant enzymes, conversion of secondary alcohols (Cn) to the corresponding primary alcohols (Cn−2) and acetic acid via ketone and ester was possible. In this way, 2-decanol was almost completely converted within 20 h at 30°C. Thus, it could be shown that the three enzymes are metabolically connected and that they are most probably involved in alkane degradation via sub-terminal oxidation of the acyclic aliphatic hydrocarbons.

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Acknowledgment

We thank the Fonds der Chemischen Industrie (Frankfurt, Germany) and the Studienstiftung des deutschen Volkes (Bonn, Germany) for stipends to Anett Kirschner.

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Authors and Affiliations

  1. Department of Biotechnology and Enzyme Catalysis, Institute of Biochemistry, Greifswald University, Felix-Hausdorff-Str. 4, 17487, Greifswald, Germany

    Anett Kirschner & Uwe T. Bornscheuer

  2. Institute of Industrial Genetics, Stuttgart University, Allmandring 31, 70569, Stuttgart, Germany

    Josef Altenbuchner

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  1. Anett Kirschner
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  2. Josef Altenbuchner
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  3. Uwe T. Bornscheuer
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Correspondence to Uwe T. Bornscheuer.

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Kirschner, A., Altenbuchner, J. & Bornscheuer, U.T. Design of a secondary alcohol degradation pathway from Pseudomonas fluorescens DSM 50106 in an engineered Escherichia coli . Appl Microbiol Biotechnol 75, 1095–1101 (2007). https://doi.org/10.1007/s00253-007-0902-3

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  • DOI: https://doi.org/10.1007/s00253-007-0902-3

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